LOCUS       sequence-406386-        5675 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  in vitro transcription of RNA encoding human ACVR1 with mutations 
            S192A and S194A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5675)
  AUTHORS   Allen RS, Jones WD, Hale M, Warder BN, Shore EM, Mullins MC
  TITLE     Reduced GS domain serine/threonine requirements of Fibrodysplasia 
            Ossificans Progressiva mutant type I BMP receptor ACVR1 in the 
            zebrafish.
  JOURNAL   J Bone Miner Res. 2023 Jun 17. doi: 10.1002/jbmr.4869.
  PUBMED    37329499
REFERENCE   2  (bases 1 to 5675)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Bone 
            Miner Res. 2023 Jun 17. doi: 10.1002/jbmr.4869."
COMMENT     Sequence Label: ACVR1-TSAA
FEATURES             Location/Qualifiers
     source          1..5675
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             1630..1677
                     /codon_start=1
                     /product="two tandem FLAG(R) epitope tags"
                     /label=2xFLAG
                     /translation="DYKDDDDKDYKDDDDK"
     polyA_signal    1727..1861
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1751..1770)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1805..1824
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1974..1994)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1974..1992)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2013..2029)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2013..2029)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2026..2048)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2037..2053
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(2061..2067,2068..2085,2086..2091))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2106..2127
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2244..2261)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2415..3003)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2495..2514)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3174..3965,3966..4034))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3797..3816
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4035..4139)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4165..4620)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4302..4323)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4514..4533
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4691..5675
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atccatggta gatggagtga tgattcttcc tgtgcttatc
      121 atgattgctc tcccctcccc tagtatggaa gatgagaagc ccaaggtcaa ccccaaactc
      181 tacatgtgtg tgtgtgaagg tctctcctgc ggtaatgagg accactgtga aggccagcag
      241 tgcttttcct cactgagcat caacgatggc ttccacgtct accagaaagg ctgcttccag
      301 gtttatgagc agggaaagat gacctgtaag accccgccgt cccctggcca agctgtggag
      361 tgctgccaag gggactggtg taacaggaac atcacggccc agctgcccac taaaggaaaa
      421 tccttccctg gaacacagaa tttccacttg gaggttggcc tcattattct ctctgtagtg
      481 ttcgcagtat gtcttttagc ctgcctgctg ggagttgctc tccgaaaatt taaaaggcgc
      541 aaccaagaac gcctcaatcc ccgagacgtg gagtatggca ctatcgaagg gctcatcacc
      601 accaatgttg gagacagcac tttagcagat ttattggatc attcgtgtac atcaggagct
      661 ggcgctggtc ttccttttct ggtacaaaga acagtggctc gccagattac actgttggag
      721 tgtgtcggga aaggcaggta tggtgaggtg tggaggggca gctggcaagg ggaaaatgtt
      781 gccgtgaaga tcttctcctc ccgtgatgag aagtcatggt tcagggaaac ggaattgtac
      841 aacactgtga tgctgaggca tgaaaatatc ttaggtttca ttgcttcaga catgacatca
      901 agacactcca gtacccagct gtggttaatt acacattatc atgaaatggg atcgttgtac
      961 gactatcttc agcttactac tctggataca gttagctgcc ttcgaatagt gctgtccata
     1021 gctagtggtc ttgcacattt gcacatagag atatttggga cccaagggaa accagccatt
     1081 gcccatcgag atttaaagag caaaaatatt ctggttaaga agaatggaca gtgttgcata
     1141 gcagatttgg gcctggcagt catgcattcc cagagcacca atcagcttga tgtggggaac
     1201 aatccccgtg tgggcaccaa gcgctacatg gcccccgaag ttctagatga aaccatccag
     1261 gtggattgtt tcgattctta taaaagggtc gatatttggg cctttggact tgttttgtgg
     1321 gaagtggcca ggcggatggt gagcaatggt atagtggagg attacaagcc accgttctac
     1381 gatgtggttc ccaatgaccc aagttttgaa gatatgagga aggtagtctg tgtggatcaa
     1441 caaaggccaa acatacccaa cagatggttc tcagacccga cattaacctc tctggccaag
     1501 ctaatgaaag aatgctggta tcaaaatcca tccgcaagac tcacagcact gcgtatcaaa
     1561 aagactttga ccaaaattga taattccctc gacaaattga aaactgactg taatcgattt
     1621 aaagctatgg attacaagga tgatgatgat aaggattaca aggatgatga tgataagaat
     1681 tcaaggcctc tcgagcctct agaactatag tgagtcgtat tacgtagatc cagacatgat
     1741 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     1801 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     1861 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt
     1921 ttaattcgcg gccgcggcgc caatgcattg ggcccggtac ccagcttttg ttccctttag
     1981 tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     2041 tatccgctca caattccaca caacatacga gccgggagca taaagtgtaa agcctggggt
     2101 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     2161 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     2221 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     2281 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     2341 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     2401 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     2461 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     2521 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     2581 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     2641 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     2701 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     2761 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     2821 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     2881 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     2941 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     3001 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     3061 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     3121 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     3181 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     3241 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     3301 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     3361 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     3421 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     3481 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     3541 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     3601 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     3661 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     3721 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     3781 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     3841 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     3901 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     3961 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     4021 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     4081 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     4141 acatttcccc gaaaagtgcc acctaaattg taagcgttaa tattttgtta aaattcgcgt
     4201 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     4261 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     4321 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     4381 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     4441 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     4501 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     4561 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     4621 cccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
     4681 tattacgcca gtcgaccgcc aattcaatat ggcgtatatg gactcatgcc aattcaatat
     4741 ggtggatctg gacctgtgcc aattcaatat ggcgtatatg gactcgtgcc aattcaatat
     4801 ggtggatctg gaccccagcc aattcaatat ggcggacttg gcaccatgcc aattcaatat
     4861 ggcggacctg gcactgtgcc aactggggag gggtctactt ggcacggtgc caagtttgag
     4921 gaggggtctt ggccctgtgc caagtccgcc atattgaatt ggcatggtgc caataatggc
     4981 ggccatattg gctatatgcc aggatcaata tataggcaat atccaatatg gccctatgcc
     5041 aatatggcta ttggccaggt tcaatactat gtattggccc tatgccatat agtattccat
     5101 atatgggttt tcctattgac gtagatagcc cctcccaatg ggcggtccca tataccatat
     5161 atggggcttc ctaataccgc ccatagccac tcccccattg acgtcaatgg tctctatata
     5221 tggtctttcc tattgacgtc atatgggcgg tcctattgac gtatatggcg cctcccccat
     5281 tgacgtcaat tacggtaaat ggcccgcctg gctcaatgcc cattgacgtc aataggacca
     5341 cccaccattg acgtcaatgg gatggctcat tgcccattca tatccgttct cacgccccct
     5401 attgacgtca atgacggtaa atggcccact tggcagtaca tcaatatcta ttaatagtaa
     5461 cttggcaagt acattactat tggaagtacg ccagggtaca ttggcagtac tcccattgac
     5521 gtcaatggcg gtaaatggcc cgcgatggct gccaagtaca tccccattga cgtcaatggg
     5581 gaggggcaat gacgcaaatg ggcgttccat tgacgtaaat gggcggtagg cgtgcctaat
     5641 gggaggtcta tataagcaat gctcgtttag ggaac
//
